Project description:RBP9-TAP was expressed in bloodstream T. brucei and bound mRNA was pulled down using IgG beads and eluted via TEV-protease cleavage. rRNA was depleted using RNAseH and rRNA hybridizing oligos.
Project description:Trypanosoma brucei were isolated from cattle, in Bunya, Uganda (Tb065BAPC) or Apuru, Uganda (Tb098AAPC). Parasites were stored as stabilities after a single mouse passage. After a further mouse passage the parasites were grown in rats to the parasitaemias indicated, isolated on DEAE cellulose and RNA was prepared. We also compared rRNA depletion with poly(A) selection.
Project description:T. brucei PF cells were treated with several chemical reagents and anti-trypanosomatid drugs. The effect of each chemical perturbation on the transcriptome of T. brucei was examined by transcript profiling of treated vs. control cells. The results indicated widespread changes, suggesting that the transcriptome of T. brucei is highly responsive to environmental factors that perturb its metabolic and biological pathways.
Project description:Trypanosoma brucei gambiense is the causative agent of the fatal human disease African sleeping sickness. Using Digital Gene Expression we have compared the transcriptome of a group 1 T.b.gambiense (Eliane) and a T.b.brucei (STIB 247).
Project description:Benzoxaboroles (BoBs) feature a boron-heterocyclic core and are an important recent innovation in the development of drugs against a range of pathogens and other pathologies. A broad spectrum of pharmacology is associated with chemically diverse BoB derivatives and includes multiple modes-of-action and targets. However, a consensus MoA for BoBs targeting evolutionarily diverse protozoan pathogens has emerged with the identification of CPSF3/CPSF73 in the CPSF complex in both apicomplexan and kinetoplastida parasites. We have detected a functional connection between protein sumoylation and the BoB boron-heterocyclic scaffold using comprehensive genetic screens in Trypanosoma brucei. Strikingly, as part of this sumoylation response, members of the CPSF complex are specifically and rapidly destabilised in a SUMO and proteosome-dependent manner. Here we deposit RNAseq data quantifying the effects of the aminomethyl-benzoxaborole AN3057 exposure on the transcriptome landscape in T. brucei. Specifically, T. brucei bloodstream-form cells in logarithmic growth phase were treated with 400 nm AN3057 (3 × EC50 determined after 24h) for 20 min (T20) and 60 min (T60), respectively. Nontreated control cells were prepared in parallel. All samples were in 2 biological replicates.
